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To assess the ecological health of the Chongming Dongtan Nature Reserve (CDNR), univariate and multimetric indices—AZTI's Marine Biotic Index (AMBI), multivariate AMBI (M-AMBI), abundance-biomass comparison curves, taxonomic diversity, Shannon-Wiener index (H′) (log2), and Benthic Opportunistic Polychaetes Amphipods index (BOPA) — were used to translate the macrobenthic community into classifications of ecological health in 2016 and 2017. Based on the results of the various indicators, the overall habitat status of the CDNR ranged from good to high status. Sites 4 and 5, which fell in the remaining tidal flats following reclamation activities, were classified as disturbed, whereas the degree of disturbance at site 3 was low. The results indicate that the CDNR is under some form of human disturbance, which includes reclamation and animal (mainly cattle) grazing. H′ (log2) and M-AMBI index may be more suitable for assessing ecological quality in intertidal zones, including the CDNR.

The water flea Daphnia pulex, a small crustacean that lives in freshwater ponds, undergoes parthenogenesis and gamogenesis according to environmental conditions. In D. pulex, different morphological characteristics can be observed using a microscope during aging. In this study, we recorded the growth, reproduction, and survival indicators of parthenogenetic D. pulex females to analyze the morphological changes that occur during senescence. We observed the growth and development of the parthenogenetic females on the 7th, 14th, 21st, and 28th days by using scanning electron microscopy and ultrastructure of the gonads on the 7th, 14th and 21st days with transmission electron microscopy. We found that body length and reproductive capacity in the parthenogenetic individuals first increased and then decreased with aging, and the survival rate decreased gradually. The depression of the brood pouch on the back of the parthenogenetic females implied a decline in gonadal function and gradual aging. During senescence, the number of lipid droplets in the gonads of the parthenogenetic females decreased, and the integrity of the mitochondria and peroxisome was destroyed. The results of our study revealed the morphological characteristics of senescence in D. pulex.

Na+/K+-ATPase (NKA) and carbonic anhydrase (CA) are ion transporters that play important roles in osmotic pressure balance in crustaceans. Oriental river prawn (Macrobrachium nipponense) is one of the most economically important shrimps in China. This species can live in freshwater and low-salt water, but the molecular mechanism of salinity regulation is unclear. In this study, full-length cDNAs of M. nipponense α-NKA and CA were obtained by rapid amplification of cDNA ends (RACE)-PCR (GenBank accession numbers MH378774 and MH827971, respectively), and characterised. Full-length cDNAs of Mnα-NKA (3778 bp) and MnCA (1407 bp) contain open reading frames of 3030 bp and 930 bp, respectively, encoding polypeptides of 1009 and 930 amino acids. Bioinformatic analysis showed that Mnα-NKA and MnCA harbour the representative features of NKAs and CAs, and share high homology with orthologs in other crustaceans. Quantitative real-time PCR revealed that Mnα-NKA and MnCA were expressed in various tissues, with highest expression in posterior gills and hepatopancreas. Both Mnα-NKA and MnCA were up-regulated in response to salinity acclimation, but expression patterns differed. Our results provide a theoretical basis for the mechanism regulating salinity acclimation in M. nipponense.

Nanoplastics are common pollutants in aquatic environments and have attracted widespread research attention. However, few studies focus on the effects of nanoplastic exposure on energy metabolism in crustaceans. Accordingly, we exposed juvenile oriental river prawns (Macrobrachium nipponense) to different concentrations of 75-nm polystyrene nanoplastics (0, 5, 10, 20, and 40 mg/L) for 7, 14, 21, or 28 days. Thereafter, the effects of nanoplastic exposure on metabolite content, energy metabolism-related enzyme activity, and gene expression were evaluated. Our results showed that (1) with increasing nanoplastic concentration and exposure time, the survival rate decreased, while weight gain rate and molting number increased and then decreased; glycogen, triglyceride, and total cholesterol content all declined while lactic acid content increased with higher exposure to nanoplastic concentrations; (2) the activities of acetyl-CoA carboxylase (ACC), hexokinase (HK), carnitine palmitoyl transferase-1, pyruvate kinase (PK), lipase, and fatty acid synthase tended to decrease, while the activity of lactate dehydrogenase (LDH) increased. In particular, the activity of 6-phosphofructokinase exposed to 5 mg/L nanoplastics increased significantly (P

Salinity is a fundamental environmental factor in aquaculture, and arginine kinase (AK) plays imperative roles in innate immune feedback and stress resistance in invertebrates. In the current study, we cloned a full-length cDNA of arginine kinase 2 (MnAK2, GenBank number, MN149533) in Macrobrachium nipponense and analyzed its function through a salinity challenge using bioinformatic approaches. MnAK2 was expressed at the highest levels in hepatopancreas and muscle. Changes in the expression levels of MnAK2, enzymes involved in innate immunity, antioxidant enzymes, and antioxidant enzyme-related genes, and the glutathione and malondialdehyde contents were investigated after 6-week salinity treatment. The expression of MnAK2 gradually increased as salinity increased, and western blotting showed that MnAK2 was significantly upregulated in the 14 and 22 ppt salinity-treatment groups relative to the control group. The findings indicate that high salinity produces oxidative stress and that salinity below isotonic salinity might improve the antioxidant response in M. nipponense. MnAK2 may play a crucial role in the response to salinity stress in M. nipponense.

Due to the nature of their habitat, which contains a high level of pollutants, estuarine crabs are at great risk of exposure to contaminants such as cadmium. Thus, in this study, the effects of cadmium on lipid metabolism were investigated in estuarine crab Chiromantes dehaani. Adult female estuarine crabs were randomly exposed to 0.05, 0.1, 0.5, and 1mg/L of CdCl2 for 7, 14 and 21days, after which the lipid contents of the hepatopancreas and ovary were measured. Also, the substance contents and the activities of the enzyme in lipid digestion, lipid synthesis and lipid transport metabolism were analyzed. The results showed that the lipid contents in the hepatopancreas and ovary of the exposed crabs decreased after prolonged exposure to cadmium compared to the control. The lipase activity decreased while the activities of fatty acid synthase and acetyl coenzyme A in the hepatopancreas increased on day 7 but decreased on days 14 and 21. Moreover, the change in non-esterified fatty acid level was similar to fatty acid synthase. The level of low-density lipoprotein increased in the exposed crabs compared to the control group while the level of high-density lipoprotein and the activity of lipoprotein lipase decreased at a higher concentration of cadmium and longer exposure time. These observations suggest that cadmium decreases the lipid content by weakening the ability of digestion, transportation and synthase of lipid, thus affecting hepatopancreas and ovary indices.

In this study, macrobenthic diversity data were collected from intertidal habitats of island wetlands in Yangtze Estuary before and after reclamation. Three survey regions based on habitat features were investigated: protected region, normal region, and self-restored region. The pattern of diversity variation showed a sharp decrease in reclamation sites and an obvious increase in vegetated sites of the self-restored region before and after reclamation. A declining trend in habitat health was observed in reclamation sites, but the degree of perturbation was relatively weaker in protected region than in normal region. The vegetated site showed a better self-restoration of biodiversity than the bald site. These results suggest that reclamation may have a negative influence on biodiversity and habitat health status in the intertidal wetland. Also, there is a possibility of self-restoration in tidal flats disturbed by reclamation and the resistance effect in nature reserve may reduce the disturbances resulting from reclamation.

In this study, several macrobenthic diversity investigations were performed in Yangtze Estuary Oyster Reef, the largest artificial oyster reef in China, from 2012 to 2014. The sampling sites of the south branch showed considerably higher diversity than those of the north branch. The richness measures exhibited a significant increasing trend from low- to high-salinity zone; however, the evenness measures were typically high in the middle-salinity zone. During the past decade, the results were combined with historical data to detect the changes in macrobenthos. The variation in substrate organisms and macrobenthic diversity followed a steady trend after a major fluctuation. Redundancy analysis indicated that the water salinity and substrate factors were the main indicators that influence macrobenthic distribution. All sampling sites in the south branch were protected by a nature reserve. However, the N2 and N6 sites in the north branch were subjected to severe and mild human interventions, respectively.

Nanoplastics are widely distributed in aquatic environments, and nanoplastic pollution has become a global concern. However, few studies have evaluated the toxicity of nanoplastics to freshwater crustaceans. In this study, by adding different concentrations of nanoplastics to water, we explored the effects of nanoplastics on the survival, antioxidant activity, immune enzyme activity, and related gene expression levels in juvenile Macrobrachium nipponense. The results showed that the 96 -h half-lethal concentration of nanoplastics to juvenile shrimp was 396.391 mg/L. As the concentration of nanoplastics increased, the activities of antioxidant enzymes generally decreased, while the contents of hydrogen peroxide and lipid peroxidation products increased. The activities of non-specific immune enzymes first increased and then decreased with increasing nanoplastic concentration. The trends in the expressions of antioxidant-related genes were generally consistent with those in the activities of antioxidant enzymes. As the nanoplastic concentration increased, the expressions of immune-related genes generally increased at first and then decreased. These results indicate that low concentrations of nanoplastics (5 mg/L) may enhance the viability of juvenile shrimp, whereas high concentrations (10,20, 40 mg/L) have inhibitory and/or toxic effects. The findings provide basic information on the toxic effects of nanoplastics in juvenile shrimp.

Recently, research on the biological effects of nanoplastics has grown exponentially. However, studies on the effects of nanoplastics on freshwater organisms and the mechanisms of the biological effects of nanoplastics are limited. In this study, the content of reactive oxygen species (ROS), gene and protein expression in the MAPK-HIF-1/NFkB pathway, and antioxidant gene expressions and enzyme activities were measured in Daphnia pulex exposed to polystyrene nanoplastic. In addition, the full-length extracellular signal-regulated kinases (ERK) gene, which plays an important role in the MAPK pathway, was cloned in D. pulex, and the amino acid sequence, function domain, and phylogenetic tree were analyzed. The results show that nanoplastic caused the overproduction of ROS along with other dose-dependent effects. Low nanoplastic concentrations (0.1 and/or 0.5 mg/L) significantly increased the expressions of genes of the MAPK pathway (ERK; p38 mitogen-activated protein kinases, p38; c-Jun amino-terminal kinases, JNK; and protein kinase B, AKT), HIF-1 pathway (prolyl hydroxylasedomain, PHD; vascular endothelial growth factor, VEGF; glucose transporter, GLUT; pyruvate kinase M, PKM; hypoxia-inducible factor 1, HIF1), and CuZn superoxide dismutase (SOD) along with the activity of glutathione-S-transferase. As the nanoplastic concentration increased, these indicators were significantly suppressed. The protein expression ratio of ERK, JNK, AKT, HIF1α, and NFkBp65 (nuclear transcription factor-kB p65) as well as the phosphorylation of ERK and NFkBp65 were increased in a dose-dependent manner. The activities of other antioxidant enzymes (catalase, total SOD, and CuZn SOD) were significantly decreased upon exposure to nanoplastic. Combined with our previous work, these results suggest that polystyrene nanoplastic causes the overproduction of ROS and activates the downstream pathway, resulting in inhibited growth, development, and reproduction. The present study fosters a better understanding of the biological effects of nanoplastics on zooplankton.

In this study, we infected juvenile red claw crayfish, Cherax quadricarinatus , with white spot syndrome virus (WSSV). We used biochemistry, transmission electron microscopy (TEM) and molecular biology methods to compare the enzymatic activities of lysozyme (LSZ), phenoloxidase (PO), peroxidase (POD), and superoxide dismutase (SOD), and morphological and CqHC gene expression changes in gills and hemolymph of juvenile C. quadricarinatus . Biochemical studies showed that, in comparison with the control group, the activities of LSZ, PO, POD and SOD in group I (injected with 50 μl WSSV) were significantly decreased after 72 h ( P P C. quadricarinatus showed that many virus particles appeared in gill tissue and the mitochondrial cristae disappeared. Hemocytes had heterochromatin in the nucleus, and there was an increase in large granule cells with abnormal volumes. Our results indicate that WSSV infection can damage the ultrastructure of the gills and hemolymph in juvenile C. quadricarinatus. Real-time polymerase chain reaction showed that CqHC gene had the highest expression level in the hepatopancreas, and expression of the HC gene sharply declined as infection progressed ( P P

In this study, intertidal macrobenthic diversity in protected, disturbed, and newly formed wetlands of Yangtze estuary was assessed using PRIMER 5.2 based on species diversity and species relatedness. We observed high diversity in nature reserves and low diversity in adjacent disturbed and newly formed wetlands. These diversity data were then integrated with historical data to detect the variation in macrobenthic diversity over the past two decades. The integrated data indicated that the intertidal macrobenthic diversity sharply decreased in heavy reclamation tidal flats whereas markedly increased in non-disturbed nature reserve and newly formed wetland. Benthic health was observed with the departure degree of average taxonomic distinctness (Δ+) and variation in taxonomic distinctness (Λ+) from the simulated 95% confidence funnel. All the habitats were subjected to different levels of human interference, except Jiuduansha and Beigangbeisha. The degradation of intertidal wetland in Yangtze estuary was mainly attributed to land reclamation, overgrazing, and overfishing.